title: Using single-molecule approaches to study archaeal DNA-binding protein Alba1.
abstract: Thermophilic and hyperthermophilic archaea have one or more copies of the Alba gene, which encodes Alba, a dimeric, highly basic protein that binds cooperatively to DNA. However, the functions of Alba and how it interacts with DNA remain unclear. In this study, we have used single-molecule tethered particle motion (TPM) and optical tweezers (OT) experiments to study the interactions between DNA molecules and Alba1. When Alba1 binds to double-stranded DNA, the Brownian motion (BM) amplitude for DNA tethers increases continuously, suggesting that Alba1 binds cooperatively. The OT study confirmed that a 5-fold increase in the persistence length of the Alba1 nucleoprotein filament is the major factor causing the increase in the BM amplitude for DNA tethers, while the contour length remained mostly unchanged. Moreover, the rate of the increase in the BM amplitude and the BM plateau value are both DNA length-dependent, indicating that the number of Alba1 initiation binding sites increases as the DNA becomes longer. Using the incoming-strand TPM experiment to monitor the interaction between Alba1 nucleoprotein filaments, we found that significant dimer-dimer contacts between two Alba1 nucleoprotein filaments are present, and the interaction is regulated by the concentration of Alba1.
authors.affiliation: Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University , 112 Taiwan.
authors.forename: Yen-Wen
authors.initials: YW
authors.lastname: Lu
authors.forename: Tao
authors.initials: T
authors.lastname: Huang
authors.forename: Cheng-Ting
authors.initials: CT
authors.lastname: Tsai
authors.forename: Yu-Yung
authors.initials: YY
authors.lastname: Chang
authors.forename: Hung-Wen
authors.initials: HW
authors.lastname: Li
authors.forename: Chun-Hua
authors.initials: CH
authors.lastname: Hsu
authors.forename: Hsiu-Fang
authors.initials: HF
authors.lastname: Fan
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D009698:Nucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D052898:Optical Tweezers; D016944:Sulfolobus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; D009698:Nucleoproteins; D004247:DNA